of cincinnati



Reissued May 31, 1927.

UNITED STA ES rn'rnu'r orrica,

CLARENCE A..MILLS, F CINCINNATZ. OHIO, ALSfiIGlN'QR T80 THE VIlLLIAM S. MEERELL COMYANY, OF CINCINNATI, GIL-10, A GGRIFORATIOEZ 0]? OHZO,

BLOOD-Ci'JAGULATISIE KEYED-YUM.

Mr invention relates to Compounds en1- pioyed in tho ins-toning of coagulation oi" blooil.

It is :1 known fact that n sultsolutinn e2;-

tract mmlr from brain tissue will hasten the coagulation of blood, although the renson for this nation not been correctly uiilerstooil uecoriling to results at my in Vestigntionu.

Tissue as; usetl in the past ha been orclinnrily brain tissue. although 1 have touncl that other tissues of the bod such as from the lung aml lziFney are richer than brain tissue in the p routs which cause blood to couguuttn fouml to l the sireil for my n; i nrouml ten ix-r cent tale, ii'tfteen per .cont proteins, nrouinl one per cent mineral salts and tho bale u-c waiter, All of the "into are not f--1)llilili3,ill stilt. solution, anti only it (GP tnio portion, of the hits combine/ll with the ,l roteins ill eliseolrr in salt mm. The nbox'e pore-grunge iiii:\l)' ilris 1ii ::'el v:1pj[)1'01\'i mute.

1 will iii tlLSL'i'llR the, es'e'olinl product of the human tissue which according to my inwhtigntions rnur e the blood to coagulate .zu'coriling to PlQSQIlt terminology this proilm-t i; n rombimition of M protein uml u phoqiliolipiu. Al. 1)lit):41)li()-lil)lll is :1 borlr 'lut iomul in the lissuos oi the lJU'lY and ronluii'iiut; phonph 1H nml the protein um. lat rompouml i:- of the class rnllod ulol ulin ilobulins :tre ('lizll'ritl(liZill by bein solu le only in (liluie salt solution. (0.5),4; Nntl.) :uui. by 1)L(%Ui|'ili21bil1l' from Sill'll :1 so llll i lz ll h'lli saturation thereof with NnCl, Nannie) llgii ll or hull saturation with (NH LSU y l have i'ountl that the epecinl ghibulin 'liiiYillg the coagulating action will sqmrnte out as n ]1I'(-"l13iliil'@ by making the solution very wool; in arid, the best nriility being .002 normzil ill'lil. Sinre this globulin is not soluble in pure water, it is possible to wnli :ill of the nrirl oil' by pure water. after \rhirh llioglobulin ru i be lissolrerl in .924 Suit so lution lo the (ll'Sli'Uil strength, nml employed :w u rougulnnt for blooil. Around of the globulin is PllOSlilKl-lllllll nml. by the u lilition of more ol' the PlIORPllO-lllilli the :i-whily ol' thrgiobulin lllt'l'ltlSl,

A; n limiter olf coimucrciul importunco Ife'o. 379,554, filed May 6, 1920. {Serial Ho. 6%,640. 4

hardly be practical n 1 n rongulont since it res. l a: l hing treatments to it since an) lnl a iitep might rlezzl which will no or 2 he tiesrroi'eil s. ill oz:-

this qlohu product m quire i obtain i in quite 31' ("Ml con ulnnt, h will pron er! to describe. rem! oi my {3 1h the bore ate at r nonstiie product I am aiuri around 26 times toe Slil'tfilfili! or solution of brain timle such of :1 sub s as has been employed in the pilot for the u)- blood to; the olil salt Water brain tissue solution.

in tho production of my conmiercinl mnterizzl i take preferably lung tissue. and after mmhimg' it; I rriml it into a salt wuier of nrouml viii/2' suit. This removes the. nrrious proteins from the tissue together with the this i are contained in (be proteins. (ert oil. nml to .rnte the solution of tissue proteins uml soiulile lnts from the remaining t-i Lie 1 hnre found. it best. to employ n 4" 'ntrilugnl machine. similar to a ('renm separator, the process being known as centru ringing.

I take other tissue, preferably brain tissuru which is rich in fats, and separate the lots from tho tissue by means of treatment with ether, alcohol. benzene and allierl produrts. carbon bisullitl, chloroform. rurbon tetra eliloiirl or the like and evaporate oil the olvent at tho lowest prnrtionl temperature. i v

l luivo then two mnterinlo. one u suit so lution of proteins. of various kinds, with my special globulin present among them nml eontainingi its; normal fut content, but with the total fat; content of the'solution wellb lilo , of the salt solution will determ amount of fats that will enter into S0l11tl0I1\ precipitating final products but I prefer that low that of normal tissue, due to the com paratively low solubility of the fats. The other material is a fatty mass containing various fats of animal tissue free of proieins.

My next step is to work the fats into the solution, which I do in some desired form of grinding mortar, in which it is possible to crush and grind. the fats in the presence of the protein solution.

All of the fats will not be absorbed in the solution, and there will be a semi-solid residue from this last operation. It should be continued, however, to an ctxtent determined by test as the point whcrecontinued addition of fats gives no strengthenin of the resultant solution; I can give no definite proportions because the richness in rdtetifis ne e therewith and because it is comparatively easy to test the action by a samp e applied toa few dro s of blood. I have found that a saturated point is reached beyond which further additions of fat are not of any value.

I have in practice added caustic soda to the final solution of the above process, to prevent a precipitation which takes place of some of the fats preferably makmg'the solution .002 normal NaOH. This precipitation does not appear to affect the strength of the it be done, if for no other purpose, to avoid the chances of bad commercial a pearance.

I do not find t at any value is conferred by this addition of the fats on coagulat ng action, except that they seem to combine with and enrich the proteins which are of the special globulin class and the coagulating force on blood of the resultant solution is increased in the fully saturated form to a large per cent over normal. The special phospho-lipins have a slight effect on coagu: lutiou as do the proteins Without the fats, but the combination of additional fat to the special globulin is the decisive factor.

I' do not wish to base my invention upon the theories advanced above but fact that the process and product result is a very greatly increased coagulating tend-- eucy on blood over any product hitherto known to the art.

It might be that the isolated globulin, (not yet named by me) may be'possible of commercial production but this I cannot predict upon the as far as my present experiments have developed. I do know that the weaker the the .tlSSll solution is in the special globulin the weaker the coagulating action on blood and that from whatever organic form the globulin dcscribedcan be obtained, it will do the desired work and will increase in activity of the special fat.

Having thus described my invention, what I claim as new and desire to secure by Letters Patent, is

1. A blood coagulant which comprises a protein, phospho-lipiu compound having the distinctive quality over related globulins of precipitating from solution upon making of the same weakly acid.

2. A blood coagulant which comprises a protein, phosphodipin compound having the distinctive quality over related globulins of from solution upon making of the same weakly acid, combined. with additional phospho-lipin.

3..A blood coagulant which comprises a protein, phospho-lipin compound having the distinctive quality over related globulins of precipitating from. solution upon making of the same weakly acid, combined with additional phosphodipin, said substance held in salt solution.

4. A blood coagulant which comprises a protein, phospho-lipin compound having the distinctive quality over related globulins of precipitating from solution upon. making of the same weakly acid, combined with additional phospho-lipin, said substance held in salt solution, and said solution made slightly alkaline suchas approximately .002 normal NaOH.

5. A blood coagulant coin rising animal tissue or extract thereof, com ined with additional phospho-lipin so as to bring the phospho-lipin content in excess of normal.

6. A blood coagulant comprising as its principal ingredients a protein and u plies pholipin and obtained by isolating the active material from a salt solution, or an extraction, or tissue.

7. A blood coagulant in a suspended, dissolved or emulsified form and comprising as its principal ingredients a protein and a phospholipin obtained by isolating the active material from a salt solution, or an extraction, of tissue.

CLARENCE A. MILLS.-

(ill 

